A novel non-invasive prenatal sickle cell disease test for all at-risk pregnancies
Corresponding Author
Julia van Campen
Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Correspondence: Julia van Campen, Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London.
E-mail: [email protected]
Search for more papers by this authorLee Silcock
Nonacus Ltd., Birmingham Research Park, Birmingham, UK
Search for more papers by this authorShu Yau
Viapath Genetics Laboratories, Guy's Hospital, London, UK
Search for more papers by this authorYvonne Daniel
Viapath Haematological Sciences Laboratories, Guy's Hospital, London, UK
Search for more papers by this authorJoo Wook Ahn
Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Search for more papers by this authorCaroline Ogilvie
Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Department of Medical and Molecular Genetics, King's College, London, UK
Search for more papers by this authorKathy Mann
Viapath Genetics Laboratories, Guy's Hospital, London, UK
Search for more papers by this authorEugene Oteng-Ntim
Department of Women and Children's Health, King's College, London, UK
Department of Women's Services, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Search for more papers by this authorCorresponding Author
Julia van Campen
Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Correspondence: Julia van Campen, Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London.
E-mail: [email protected]
Search for more papers by this authorLee Silcock
Nonacus Ltd., Birmingham Research Park, Birmingham, UK
Search for more papers by this authorShu Yau
Viapath Genetics Laboratories, Guy's Hospital, London, UK
Search for more papers by this authorYvonne Daniel
Viapath Haematological Sciences Laboratories, Guy's Hospital, London, UK
Search for more papers by this authorJoo Wook Ahn
Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Search for more papers by this authorCaroline Ogilvie
Genetics Laboratories, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Department of Medical and Molecular Genetics, King's College, London, UK
Search for more papers by this authorKathy Mann
Viapath Genetics Laboratories, Guy's Hospital, London, UK
Search for more papers by this authorEugene Oteng-Ntim
Department of Women and Children's Health, King's College, London, UK
Department of Women's Services, Guy's and St. Thomas' NHS Foundation Trust, London, UK
Search for more papers by this authorSummary
Sickle cell disease (SCD) is the most common genetic haematological disorder. The availability of non-invasive prenatal diagnosis (NIPD) is predicted to increase uptake of prenatal diagnosis for SCD, as it has no perceived procedure-related miscarriage risk. We report the development of a targeted massively parallel sequencing (MPS) assay for the NIPD of fetal SCD using fetal cell-free (cf)DNA from maternal plasma, with no requirement for paternal or proband samples. In all, 64 plasma samples from pregnant women were analysed: 42 from SCD carriers, 15 from women with homozygous (Hb SS) SCD and seven from women with compound heterozygous (Hb SC) SCD. Our assay incorporated a relative mutation dosage assay for maternal carriers and a wild type allele detection assay for affected women (Hb SS/Hb SC). Selective analysis of only smaller cfDNA fragments and modifications to DNA fragment hybridisation capture improved diagnostic accuracy. Clinical sensitivity was 100% and clinical specificity was 100%. One sample with a fetal fraction of <4% was correctly called as ‘unaffected’, but with a discordant genotype (Hb AA rather than Hb AS). Six samples gave inconclusive results, of which two had a fetal fraction of <4%. This study demonstrates that NIPD for SCD is approaching clinical utility.
Supporting Information
| Filename | Description |
|---|---|
| bjh16529-sup-0001-Supinfo1.docxWord document, 464.8 KB |
Figure S1. Fetal fraction correlates with gestational age. Figure S2. Modified Sequential Probability Ratio Test (SPRT) curves for three samples, before and after in silico selection of fragments <155bp. Table SI. Results for all samples - RMD. Table SII. Results for all samples – wild type allele detection. |
| bjh16529-sup-0002-Supinfo2.docxWord document, 319 KB |
Data S1. Laboratory workflow with timings. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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